Application of antibody CD29 in identification and sorting of human-placenta-chorion-avillosum-derived mesenchymal stem cells

A technology of mesenchymal stem cells and human placenta, applied in animal cells, vertebrate cells, cell dissociation methods, etc., can solve the dysfunction of human placental smooth chorionic mesenchymal stem cells Stem cell difficulties and other problems, to achieve rapid and accurate identification, good detection stability

Inactive Publication Date: 2020-12-11
CHENGDU QINGKE BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This brings difficulties to the identification and sorting of qualified human placental smooth chorionic mesenchymal stem cells ...

Method used

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  • Application of antibody CD29 in identification and sorting of human-placenta-chorion-avillosum-derived mesenchymal stem cells
  • Application of antibody CD29 in identification and sorting of human-placenta-chorion-avillosum-derived mesenchymal stem cells
  • Application of antibody CD29 in identification and sorting of human-placenta-chorion-avillosum-derived mesenchymal stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: Isolation and culture of human smooth chorion-derived mesenchymal stem cells

[0029] The method for isolating and culturing human smooth chorion-derived mesenchymal stem cells comprises the following steps:

[0030] (1) Collect maternal blood before delivery for infectious disease testing, including syphilis, AIDS, hepatitis B, hepatitis C, etc., and select healthy full-term placenta;

[0031] (2) Cut the fetal membrane tissue at the edge of the placenta, cut it into an appropriate size, wash the fetal membrane tissue with tissue cleaning solution, wash away the blood, and then use tissue forceps to peel off the upper layer of amniotic membrane tissue, and then peel off the smooth chorion layer, put it in a plate for cleaning and remove residual blood vessels;

[0032] (3) Transfer the separated smooth chorionic tissue to a 50mL centrifuge tube, cut to about 1mm 3 ~3mm 3 Add tissue cleaning solution to the tissue block, and centrifuge at 700×g for 5 minut...

Embodiment 2

[0038] Example 2: Detection of PMSCs cell surface markers

[0039] Resuscitate the cryopreserved P1 generation cells in groups A and B, the number of cells should not be less than 2×10 6 After centrifugation and washing, CD11b, CD19, CD34, CD45, HLA-D, CD29, CD73, CD90, CD105 and other antibodies were used to label respectively, resuspended to 400 μl in PBS, and detected by flow cytometry. The detection results were as follows: Figure 4-11 Shown, and the pass rate of cell flow cytometry results in groups A and B were counted, and the results are listed in Table 1.

[0040] Table 1 Flow detection results

[0041]

[0042] From Figure 4-11 And it can be seen from Table 1 that the CD90 of group A is unqualified, the CD90 of group B is qualified, and the CD29 of groups A and B are both qualified. It shows that CD29 has a good detection effect in the detection of human placental smooth chorion-derived mesenchymal stem cells.

[0043] Comparing the three-way differentiation...

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Abstract

The invention discloses an application method of CD29 in identification and sorting of human-placenta-chorion-avillosum-derived mesenchymal stem cells. According to the application method disclosed bythe invention, the human-placenta-chorion-avillosum-derived mesenchymal stem cells (PMSCs) are firstly marked with a variety of cell surface markers separately, then, detection is carried out by using a flow cytometer to survey qualified rate and stability of flow-type detection marked with all the markers, and it is discovered that the expression of a marker CD90 in a universal standard is unstable, and detections are all qualified after the PMSCs are marked with the CD29; and through repeated test, it is discovered that the CD29 has good detection stability, the qualified rate of detectionis 100%, thus, the CD29 has a good and stable identification effect on the PMSCs and can be applied to rapid and accurate identification on the PMSCs instead of the CD90.

Description

technical field [0001] The invention belongs to the technical field of identification of mesenchymal stem cells. Background technique [0002] In the basic research or clinical research of biomedicine, when analyzing the cell number and function changes of a specific cell group, it is necessary to separate this group of cells from other cells. At this time, it is necessary to separate them according to different cell surface markers. separate. Cell surface markers refer to membrane proteins embedded in the lipid bilayer structure of cell membranes, including membrane antigens, membrane receptors and other molecules. CD molecules (Cluster of Differentiation, differentiation clusters or differentiation groups, also called leukocyte differentiation antigens) are One of the most commonly used cell surface markers refers to the application of monoclonal antibody identification-based methods to identify the same differentiation antigen recognized by monoclonal antibodies from dif...

Claims

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Application Information

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IPC IPC(8): C12N5/0775G01N33/569
CPCC12N5/0668G01N33/56966C12N2509/00C12N2509/10G01N2333/7055
Inventor 高雪华孙雯刘倩伶曾焰江蕊利侯爽海泉
Owner CHENGDU QINGKE BIOTECH
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